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目的:观察熟地有效成分甘露三糖对高浓度皮质酮诱导海马神经元损伤及学习记忆相关信号转导分子表达的影响。方法:用24小时新生大鼠原代培养海马神经细胞。第八天用药处理,将细胞分为5组:1:正常对照组;2:1×10-4M高浓度皮质酮模型组;3:10-4 M CORT+3nM RU38486拮抗剂组;4:10-4 M CORT+0.1g/L甘露三糖组;5:10-4 M CORT+40uM多奈哌齐阳性对照组。24h后,通过SYTO13-PI双荧光染色观察各组细胞的形态及存活情况,MTT法测定细胞活性,Western Blot检测学习记忆信号转导通路相关分子GCR、BDNF、SGK蛋白表达。结果:与正常组相比,模型组细胞死亡较多,细胞活力下降,GCR、BDNF、SGK蛋白表达均显著性降低;如上改变又均被皮质酮受体拮抗剂RU38486逆转;甘露三糖(0.1g/L)同阳性对照多奈派齐(40μM)均明显减少死细胞数量,提高细胞活力,提高模型细胞GCR、BDNF、SGK蛋白表达。结论:熟地有效成分甘露三糖保护大鼠海马神经细胞免遭高浓度皮质酮的损伤,通过调节学习记忆信号转导途径中的重要蛋白GCR、BDNF、SGK表达从而改善高浓度皮质酮致学习记忆功能退化。
OBJECTIVE: To observe the effects of Rehmannia glutinosa, an effective ingredient, on the injury of hippocampal neurons induced by high concentrations of corticosterone and the expression of signal transduction molecules involved in learning and memory. Methods: Primary cultured hippocampal neurons were cultured in neonatal rats for 24 hours. On the eighth day, the cells were divided into 5 groups: 1: normal control group, 2: 1 × 10-4M high concentration corticosterone model group, 3: 10-4 M CORT + 3nM RU38486 antagonist group, 4: 10 -4 M CORT + 0.1 g / L mannosetriose group; 5: 10-4 M CORT + 40 uM donepezil positive control group. After 24 hours, the morphology and survival of the cells were observed by SYTO13-PI double staining. The cell viability was measured by MTT assay. The expression of GCR, BDNF and SGK proteins were detected by Western Blot. Results: Compared with the normal group, the model group showed more cell death and decreased cell viability, and the expressions of GCR, BDNF and SGK were significantly lower than those in the normal group (P <0.05). The changes were also reversed by the RU38486 corticosterone receptor antagonist. g / L) and the positive control Donepaz (40μM) significantly decreased the number of dead cells, increased cell viability, and increased the expression of GCR, BDNF and SGK in model cells. Conclusion: The traditional Chinese medicinal ingredient mannotriose protects rat hippocampal neurons from high concentration corticosterone, and improves learning and memory of high concentration corticosterone by regulating the expression of important proteins in learning and memory signal transduction pathway, such as GCR, BDNF and SGK Degradation of function.